CN104865348B - A kind of method of efficient detection nano-particle myocardial toxicity - Google Patents
A kind of method of efficient detection nano-particle myocardial toxicity Download PDFInfo
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Abstract
A kind of method that the invention discloses efficient detection nano-particle myocardial toxicity, the method comprises the steps: 1) prepare patterned fibrous electrospinning reception plate, utilize optical etching technology and magnetically controlled DC sputtering technology, preparation has the reception plate of pattern metal silver layer；2) preparation patterning electrospinning fibre, utilizes step 1) gains to carry out electrostatic spinning；3) PDMS cavity is prepared；4) by step 2) and step 3) gains plasma treatment be attached；5) Primary mouse myocardial cell is inoculated on the patterning electrospinning fibre in step 4) gains, utilize syringe pump that culture medium pumps into the cavity of step 4) gains, again nano-particle is pumped into, pump in nano-particle and in latter 20 days, record various myocardial toxicity index.The present invention can realize nano-particle myocardial toxicity efficient, high-precision detection, and Detection results is consistent with interior animal experiment；More quick relative to interior animal experiment.
Description
Technical field
The invention belongs to nano-particle toxicity detection field, a kind of method being specifically related to efficient detection nano-particle myocardial toxicity.
Technical background
At biomedicine field, nano-particle is commonly used as the carrier of medicine conveying, when using nano-particle, often need to its toxicity be evaluated.
Heart is the vitals of human body, and can a kind of nano-particle carry out bio-medical, and cardiocellular toxicity is important investigation object by it.
In the prior art, the common method of the myocardial toxicity evaluating nano cell is carry out the zoopery of nano-particle toxicity.But carrying out zoopery, the required detection cycle is long, with often causing detection error due to the individual variation of laboratory animal.Prior art is used for substitute zooperal experiment in vitro, complex steps, it is easy to owing to operational error causes error, degree of accuracy is difficult to ensure that.
Summary of the invention
For the shortcoming of prior art, a kind of method that it is an object of the invention to provide efficient detection nano-particle myocardial toxicity, it is characterised in that described method comprises the steps:
1) prepare patterned fibrous electrospinning and receive plate: on insulating glass sheet, be first coated with positive-working photoresist, then cover lid layer photomask, utilize litho machine to perform etching；Again through deposition layer of metal silver on direct magnetic control technology sheet glass after etching, the shape of the argent deposited includes the one in square, rectangle, the length of side range for 20 μm-100 μm；Finally wash remaining positive-working photoresist；
2) preparation patterning electrospinning fibre: by medical high polymer organic solvent dissolution, utilize step 1) gains to receive plate as electrospinning, prepared the patterning electrospinning fibre of square or rectangular composition by electrostatic spinning technique；The diameter controlling gained fiber is 100-300nm；
Described medical high polymer includes the one in polylactic acid, polycaprolactone, polyurethane, polyacrylonitrile, and described organic solvent includes at least one in acetone, dimethylformamide；
3) negative epoxy resin type nearultraviolet rays photoresist SU-8 is placed on silicon chip, utilizes optical etching technology, remove remaining SU-8, retain width be 40-150 μm, height be that the SU-8 cuboid of 70-100 μm is as mould；Melted PDMS is placed on gained mould, after PDMS cools down, removes mould, obtain PDMS cavity；
4) by step 2) and step 3) gains at O2Or N2Atmosphere under, by Cement Composite Treated by Plasma 60 seconds so that step 2) and step 3) gains compact siro spinning technology；
5) by 5 × 106-1.0×107Individual mouse primary myocardial cell is inoculated on the patterning electrospinning fibre in step 4) gains, utilizes syringe pump that culture medium pumps into the cavity of step 4) gains, and flow velocity is 0.2-0.3ml/h；Again nanoparticle suspension is pumped into after PBS, after nano-particle pumps into, record the change of the change of mouse primary myocardial cell jumping frequency rate, the enzyme work of superoxide dismutase, the activity of glutathion peroxidase, mda content, the damage of cell DNA, apoptosis rate, draw the toxicity of nano-particle；
Described nano-particle includes the one in nano ferriferrous oxide, nano titanium oxide, nano zine oxide, nano silicon, nano-Ag particles.
The present invention gropes to find by substantial amounts of experiment, it is the electrospinning fibre of 100-300nm when selecting diameter, and when the pattern of fibrous membrane is set to square or rectangular array, and arrange PDMS cavity be of a size of wide for 40-150 μm, high for 70-100 μm time, mouse primary myocardial cell can be made well to grow in cavity, and nano-particle is produced almost identical with zoopery toxic reaction.
The present invention can so that patterned fibrous and PDMS cavity and substrate of glass is seamless fits tightly, it is ensured that nano-particle completely with cells contacting, it is ensured that the degree of accuracy of detection.Simultaneously as patterned fibrous film is fitted with substrate and cavity closely so that the present invention can have good repetitive operation, saves the cost of detection.
It is essential that any predictable change made according to the present invention, as finely tuned pattern form and PDMS size, belong to protection scope of the present invention.
Special needs to be pointed out is, the present invention is applicable to the myocardial toxicity of detection nano-particle.General knowledge according to this area, it should be understood that the present invention detects suitable in the myocardial toxicity of any nano-particle.It should be understood that several nano-particle cited by the present invention are used only for proving that the present invention detects suitable in the myocardial toxicity of nano-particle, should not be construed as limitation of the invention.
In the present invention, the argent of step 1) deposition can be prepared as any type of array, and only needing the figure in array is square or rectangular of the present invention.
Preferably, in described step 1) deposition argent be shaped as square, the length of side range for 20 μm-100 μm.When being shaped as quadrate array of the argent deposited, primary cardiomyocytes has better growth conditions, is more beneficial for the detection of the myocardial toxicity of nano-particle.
Described step 2) in, when carrying out electrospinning, voltage is 15-25KV, and flow velocity is 0.5-1.0ml/h.
Preferably, described step 2) in, the diameter of fiber is 200nm.When the diameter of fiber used is 200nm, primary cardiomyocytes has better growth conditions, is more beneficial for the detection of the myocardial toxicity of nano-particle.
Described step 2) in medical high polymer be polylactic acid, described organic solvent is the mixed solution of acetone and dimethylformamide, and the volume ratio of acetone and dimethylformamide is 9:1.When fiber is acid fiber by polylactic, primary cardiomyocytes has better growth conditions, is more beneficial for the detection of the myocardial toxicity of nano-particle.
Preferably, the wide of the mould in described step 3) is 100 μm, and height is 80 μm.
It is furthermore preferred that the wide of mould in described step 3) is 50 μm, height is 70 μm.Detection tract is more little, more integrated, it is possible to achieve a detecting device has multiple tract, it is achieved the carrying out of more groups of parallel laboratory tests, more guarantees the degree of accuracy of detection.In the prior art, not yet it is found that prepare cavity so little, carry out myocardial toxicity detection based on electrospun fiber membrane.
In described step 5), the number of the primary cardiomyocytes for inoculating is 8 × 106Individual.
In described step 5), the flow velocity of culture medium is 0.25ml/h.
Beneficial effects of the present invention:
1, the present invention can realize nano-particle myocardial toxicity efficient, high-precision detection, and Detection results is consistent with interior animal experiment；More quick relative to interior animal experiment；
2, the present invention is raw materials used is easy to get, and cost is low, and technology maturation used is easily implemented, and has huge market application foreground.
Accompanying drawing explanation
Fig. 1 is gained patterned fibrous of the present invention.
Detailed description of the invention
The foregoing of the present invention is described in further detail by detailed description of the invention by the following examples, but this should not being interpreted as, the scope of the above-mentioned theme of the present invention is only limitted to Examples below.
Embodiment 1
1) prepare patterned fibrous electrospinning and receive plate: on insulating glass sheet, be first coated with positive-working photoresist, then cover lid layer photomask, utilize litho machine to perform etching；Again through deposition layer of metal silver on direct magnetic control technology sheet glass after etching, the argent deposited be shaped as quadrate array, the length of side range for 20 μm；Finally wash remaining positive-working photoresist；
2) preparation patterning electrospinning fibre: by medical high polymer organic solvent dissolution, utilize step 1) gains to receive plate as electrospinning, prepared the patterning electrospinning fibre of quadrate array by electrostatic spinning technique；The diameter controlling gained fiber is 100nm；
Described medical high polymer is polyacrylonitrile, and described organic solvent is acetone and dimethylformamide mixed solution (9:1v/v)；
3) SU-8 is placed on silicon chip, utilizes optical etching technology, remove remaining SU-8, retain wide be 100 μm, height be that the SU-8 cuboid of 80 μm is as mould；Melted PDMS is placed on gained mould, after PDMS cools down, removes mould, obtain PDMS cavity；
4) passing through plasma treatment: open equipment vacuum room, loading good deposition prepared above has sheet glass and the PDMS cavity of patterning electrospinning fibre, opens RF driving source filament supply, opens O2:Gas cylinder valve, Cement Composite Treated by Plasma is after 1 minute, by step 2) and step 3) gains fit tightly so that it is connect；
5) by 8 × 106Individual mouse primary myocardial cell is inoculated on the patterning electrospinning fibre in step 4) gains, utilizes syringe pump that culture medium pumps into the cavity of step 4) gains, and flow velocity is 0.25ml/h；Again nanoparticle suspension is pumped into after PBS, after nano-particle pumps into, record the change of the cell jumping frequency rate change of mouse primary myocardial cell, the enzyme work of superoxide dismutase, the activity of glutathion peroxidase, mda content, the damage of cell DNA, apoptosis rate, draw the toxicity of nano-particle；
Described nano-particle includes nano-Ag particles.
Embodiment 2
1) prepare patterned fibrous electrospinning and receive plate: on insulating glass sheet, be first coated with positive-working photoresist, then cover lid layer photomask, utilize litho machine to perform etching；Again through deposition layer of metal silver on direct magnetic control technology sheet glass after etching, the argent deposited be shaped as rectangular array, the length of side is 20 μm and 70 μm, finally washes remaining positive-working photoresist；
2) preparation patterning electrospinning fibre: by medical high polymer organic solvent dissolution, utilize step 1) gains to receive plate as electrospinning, prepared the patterning electrospinning fibre of rectangular array by electrostatic spinning technique；The diameter controlling gained fiber is 150nm；
Described medical high polymer includes polylactic acid, and described organic solvent is acetone；
3) SU-8 is placed on silicon chip, utilizes optical etching technology, remove remaining SU-8, retain wide be 100 μm, height be that the SU-8 cuboid of 80 μm is as mould；Melted PDMS is placed on gained mould, after PDMS cools down, removes mould, obtain PDMS cavity；
4) passing through plasma treatment: open equipment vacuum room, loading good deposition prepared above has sheet glass and the PDMS cavity of patterning electrospinning fibre, opens RF driving source filament supply, opens N2Gas cylinder valve, Cement Composite Treated by Plasma is after 1 minute, by step 2) and step 3) gains fit tightly so that it is connect；
5) by 8 × 106Individual mouse primary myocardial cell is inoculated on the patterning electrospinning fibre in step 4) gains, utilizes syringe pump that culture medium pumps into the cavity of step 4) gains, and flow velocity is 0.25ml/h；Again nanoparticle suspension is pumped into after PBS, after nano-particle pumps into, record the change of the cell jumping frequency rate change of mouse primary myocardial cell, the enzyme work of superoxide dismutase, the activity of glutathion peroxidase, mda content, the damage of cell DNA, apoptosis rate, draw the toxicity of nano-particle；
Described nano-particle includes nano silicon.
Embodiment 3
1) prepare patterned fibrous electrospinning and receive plate: on insulating glass sheet, be first coated with positive-working photoresist, then cover lid layer photomask, utilize litho machine to perform etching；Again through deposition layer of metal silver on direct magnetic control technology sheet glass after etching, the argent deposited be shaped as quadrate array, the length of side is 50 μm；Finally wash remaining positive-working photoresist；
2) preparation patterning electrospinning fibre: by medical high polymer organic solvent dissolution, utilize step 1) gains to receive plate as electrospinning, prepare square pattern electrospinning fibre by electrostatic spinning technique；The diameter controlling gained fiber is 200nm；
Described medical high polymer includes polyurethane, and described organic solvent is acetone；
3) SU-8 is placed on silicon chip, utilizes optical etching technology, remove remaining SU-8, retain wide be 100 μm, height be that the SU-8 cuboid of 80 μm is as mould；Melted PDMS is placed on gained mould, after PDMS cools down, removes mould, obtain PDMS cavity；
4) passing through plasma treatment: open equipment vacuum room, loading good deposition prepared above has sheet glass and the PDMS cavity of patterning electrospinning fibre, opens RF driving source filament supply, opens N2Gas cylinder valve, Cement Composite Treated by Plasma is after 1 minute, by step 2) and step 3) gains fit tightly so that it is connect；
5) by 8 × 106Individual mouse primary myocardial cell is inoculated on the patterning electrospinning fibre in step 4) gains, utilizes syringe pump that culture medium pumps into the cavity of step 4) gains, and flow velocity is 0.2-0.3ml/h；Again nanoparticle suspension is pumped into after PBS, after nano-particle pumps into, record the change of the cell jumping frequency rate change of mouse primary myocardial cell, the enzyme work of superoxide dismutase, the activity of glutathion peroxidase, mda content, the damage of cell DNA, apoptosis rate, draw the toxicity of nano-particle；
Described nano-particle is nano zine oxide.
Embodiment 4
1) prepare patterned fibrous electrospinning and receive plate: on insulating glass sheet, be first coated with positive-working photoresist, then cover lid layer photomask, utilize litho machine to perform etching；Again through deposition layer of metal silver on direct magnetic control technology sheet glass after etching, the argent deposited be shaped as rectangular array, the length of side respectively 50 μm and 75 μm；Finally wash remaining positive-working photoresist；
2) preparation patterning electrospinning fibre: by medical high polymer organic solvent dissolution, utilize step 1) gains to receive plate as electrospinning, by electrostatic spinning technique preparation patterning electrospinning fibre；The diameter controlling gained fiber is 250nm；
Described medical high polymer is polylactic acid, and described organic solvent is acetone；
3) SU-8 is placed on silicon chip, utilizes optical etching technology, remove remaining SU-8, retain wide be 150 μm, height be that the SU-8 cuboid of 100 μm is as mould；Melted PDMS is placed on gained mould, after PDMS cools down, removes mould, obtain PDMS cavity；
4) passing through plasma treatment: open equipment vacuum room, loading good deposition prepared above has sheet glass and the PDMS cavity of patterning electrospinning fibre, opens RF driving source filament supply, opens O2Gas cylinder valve, Cement Composite Treated by Plasma is after 1 minute, by step 2) and step 3) gains fit tightly so that it is connect；
5) by 1.0 × 107Individual mouse primary myocardial cell is inoculated on the patterning electrospinning fibre in step 4) gains, utilizes syringe pump that culture medium pumps into the cavity of step 4) gains, and flow velocity is 0.3ml/h；Again nanoparticle suspension is pumped into after PBS, after nano-particle pumps into, record the change of the cell jumping frequency rate change of mouse primary myocardial cell, the enzyme work of superoxide dismutase, the activity of glutathion peroxidase, mda content, the damage of cell DNA, apoptosis rate, draw the toxicity of nano-particle；
Described nano-particle is nano ferriferrous oxide.
Embodiment 5
1) prepare patterned fibrous electrospinning and receive plate: on insulating glass sheet, be first coated with positive-working photoresist, then cover lid layer photomask, utilize litho machine to perform etching；Again through deposition layer of metal silver on direct magnetic control technology sheet glass after etching, the argent deposited be shaped as rectangular array, the length of side range for 20 μm and 100 μm；Finally wash remaining positive-working photoresist；
2) preparation patterning electrospinning fibre: by medical high polymer organic solvent dissolution, utilize step 1) gains to receive plate as electrospinning, by electrostatic spinning technique preparation patterning electrospinning fibre；The diameter controlling gained fiber is 300nm；
Described medical high polymer is polycaprolactone, and described organic solvent is acetone；
3) SU-8 is placed on silicon chip, utilizes optical etching technology, remove remaining SU-8, retain wide be 40 μm, height be that the SU-8 cuboid of 70 μm is as mould；Melted PDMS is placed on gained mould, after PDMS cools down, removes mould, obtain PDMS cavity；
4) passing through plasma treatment: open equipment vacuum room, loading good deposition prepared above has sheet glass and the PDMS cavity of patterning electrospinning fibre, opens RF driving source filament supply, opens O2Gas cylinder valve, Cement Composite Treated by Plasma is after 1 minute, by step 2) and step 3) gains fit tightly so that it is connect；
5) by 5 × 106Individual mouse primary myocardial cell is inoculated on the patterning electrospinning fibre in step 4) gains, utilizes syringe pump that culture medium pumps into the cavity of step 4) gains, and flow velocity is 0.2ml/h；Again nanoparticle suspension is pumped into after PBS, after nano-particle pumps into, record the change of the cell jumping frequency rate change of mouse primary myocardial cell, the enzyme work of superoxide dismutase, the activity of glutathion peroxidase, mda content, the damage of cell DNA, apoptosis rate, draw the toxicity of nano-particle；
Described nano-particle is nano titanium oxide.
Experimental example 1:
The method utilizing the embodiment of the present invention 1 carries out myocardial toxicity detection: when Primary mouse myocardial cell is in system of the present invention after In vitro culture 20 days, adding concentration is 80 μ g/ml, when particle diameter is about the nano-Ag particles of 80nm, after acting on 24 hours, the jumping frequency rate recording primary cardiomyocytes is 243 beats/min, the activity measuring its superoxide dismutase (SOD) is 10.44U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 47.14 μm of ol/gprot, the content of malonaldehyde (MDA) is 1.45nmol/mgprot, the damage ratio of DNA is 8.32%, apoptosis rate is 6.74%.
Experimental example 2:
The method utilizing the embodiment of the present invention 1 carries out myocardial toxicity detection: when Primary mouse myocardial cell is in system of the present invention after In vitro culture 15 days, adding concentration is 60 μ g/ml, when particle diameter is about the nano silicon of 70nm, after acting on 24 hours, the jumping frequency rate recording primary cardiomyocytes is 174 beats/min, the activity measuring its superoxide dismutase (SOD) is 35.56U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 54.03 μm of ol/gprot, the content of malonaldehyde (MDA) is 10.61nmol/mgprot, the damage ratio of DNA is 56.11%, apoptosis rate is 46.88%.
Experimental example 3:
The method utilizing the embodiment of the present invention 1 carries out myocardial toxicity detection: when Primary mouse myocardial cell is in system of the present invention after In vitro culture 10 days, adding concentration is 100 μ g/ml, when particle diameter is about the nano zine oxide of 50nm, after acting on 24 hours, the jumping frequency rate recording primary cardiomyocytes is 168 beats/min, the activity measuring its superoxide dismutase (SOD) is 54.32U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 67.35 μm of ol/gprot, the content of malonaldehyde (MDA) is 18.97nmol/mgprot, the damage ratio of DNA is 68.15%, apoptosis rate is 60.04%.
Experimental example 4:
The method utilizing the embodiment of the present invention 1 carries out myocardial toxicity detection: when Primary mouse myocardial cell is in system of the present invention after In vitro culture 10 days, adding concentration is 400 μ g/ml, when particle diameter is about the ferroso-ferric oxide of 30nm, after acting on 24 hours, the jumping frequency rate recording primary cardiomyocytes is 113 beats/min, the activity measuring its superoxide dismutase (SOD) is 87.46U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 79.57U/mgprot, the content of malonaldehyde (MDA) is 45.33nmol/mgprot, the damage ratio of DNA is 78.43%, apoptosis rate is 72.51%.
Experimental example 5:
The method utilizing the embodiment of the present invention 1 carries out myocardial toxicity detection: when Primary mouse myocardial cell is in system of the present invention after In vitro culture 10 days, adding concentration is 70 μ g/ml, when particle diameter is about the titanium dioxide of 10nm, after acting on 24 hours, the jumping frequency rate recording primary cardiomyocytes is 207 beats/min, the activity measuring its superoxide dismutase (SOD) is 43.15U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 13.28U/mgprot, the content of malonaldehyde (MDA) is 8.57nmol/mgprot, the damage ratio of DNA is 28.67%, apoptosis rate is 22.74%.
Contrast experiment's example 1:
Mice carried out tail vein injection experiment, implantation concentration is 80 μ g/ml, particle diameter is about the nanometer silver of 80nm and is about 5ml, effect body is after 24 hours, recording mouse heart jumping frequency rate is 266 beats/min, take out mouse heart, carry out homogenate detection, the activity measuring the superoxide dismutase (SOD) in heart is 15.65U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 49.58U/mgprot, the damage ratio that content is 2.37nmol/mgprot, DNA of malonaldehyde (MDA) is 3.58%, and apoptosis rate is 3.44%.
Contrast experiment's example 2:
Mice carried out tail vein injection experiment, implantation concentration is 60 μ g/ml, particle diameter is about the nano silicon of 70nm and is about 5ml, effect body is after 24 hours, recording mouse heart jumping frequency rate is 198 beats/min, take out mouse heart, carry out homogenate detection, the activity measuring the superoxide dismutase (SOD) in heart is 37.89U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 59.74U/mgprot, the damage ratio that content is 12.54nmol/mgprot, DNA of malonaldehyde (MDA) is 50.33%, and apoptosis rate is 40.24%.
Contrast experiment's example 3:
Mice carried out tail vein injection experiment, implantation concentration is 100 μ g/ml, particle diameter is about the nano zine oxide of 50nm and is about 5ml, effect body is after 24 hours, recording mouse heart jumping frequency rate is 185 beats/min, take out mouse heart, carry out homogenate detection, the activity measuring the superoxide dismutase (SOD) in heart is 57.67U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 70.25U/mgprot, the damage ratio that content is 21.07nmol/mgprot, DNA of malonaldehyde (MDA) is 59.18%, and apoptosis rate is 52.66%.
Contrast experiment's example 4:
Mice carried out tail vein injection experiment, implantation concentration is 400 μ g/ml, particle diameter is about the nano zine oxide of 30nm and is about 5ml, effect body is after 24 hours, recording mouse heart jumping frequency rate is 142 beats/min, take out mouse heart, carry out homogenate detection, the activity measuring the superoxide dismutase (SOD) in heart is 89.17U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 80.12U/mgprot, the damage ratio that content is 47.33nmol/mgprot, DNA of malonaldehyde (MDA) is 69.54%, and apoptosis rate is 65.37%.
Contrast experiment's example 5:
Mice carried out tail vein injection experiment, implantation concentration is 70 μ g/ml, particle diameter is about the nano titanium oxide of 10nm and is about 5ml, effect body is after 24 hours, recording mouse heart jumping frequency rate is 232 beats/min, take out mouse heart, carry out homogenate detection, the activity measuring the superoxide dismutase (SOD) in heart is 47.54U/mgprot, the activity of glutathion peroxidase (GSH-Px) is 16.35U/mgprot, the damage ratio that content is 9.67nmol/mgprot, DNA of malonaldehyde (MDA) is 21.93%, and apoptosis rate is 20.07%.
By above-mentioned experimental example 1-5 and contrast experiment example 1-5 it can be seen that in the scheming toxicity detection of nano-particle, the testing result of the present invention is similar to interior animal experiment result, can as the replacement scheme of interior animal experiment detection.It is essential that the present invention is more easy to control than experiment in vivo, and Financial cost and time cost lower, there is very good market application foreground.
Claims (9)
1. the method for an efficient detection nano-particle myocardial toxicity, it is characterised in that described method comprises the steps:
1) prepare patterned fibrous electrospinning and receive plate: on insulating glass sheet, be first coated with positive-working photoresist, then cover lid layer photomask, utilize litho machine to perform etching；Again through deposition layer of metal silver on direct magnetic control technology sheet glass after etching, the shape of the argent deposited includes the one in square, rectangle, the length of side range for 20 μm-100 μm；Finally wash remaining positive-working photoresist；
2) preparation patterning electrospinning fibre: by medical high polymer organic solvent dissolution, utilize step 1) gains to receive plate as electrospinning, prepared the patterning electrospinning fibre of square or rectangular composition by electrostatic spinning technique；The diameter controlling gained fiber is 100-300nm；
Described medical high polymer includes the one in polylactic acid, polycaprolactone, polyurethane, polyacrylonitrile, and described organic solvent includes at least one in acetone, dimethylformamide；
3) negative epoxy resin type nearultraviolet rays photoresist SU-8 is placed on silicon chip, utilizes optical etching technology, remove remaining SU-8, retain width be 40-150 μm, height be that the SU-8 cuboid of 70-100 μm is as mould；Melted PDMS is placed on gained mould, after PDMS cools down, removes mould, obtain PDMS cavity；
4) by step 2) and step 3) gains at O2Or N2Atmosphere under, by Cement Composite Treated by Plasma 60 seconds so that step 2) and step 3) gains compact siro spinning technology；
5) by 5 × 106-1.0×107Individual mouse primary myocardial cell is inoculated on the patterning electrospinning fibre in step 4) gains, utilizes syringe pump that culture medium pumps into the cavity of step 4) gains, and flow velocity is 0.2-0.3ml/h；Again nanoparticle suspension is pumped into after PBS, after nano-particle pumps into, record the change of the change of mouse primary myocardial cell jumping frequency rate, the enzyme work of superoxide dismutase, the activity of glutathion peroxidase, mda content, the damage of cell DNA, apoptosis rate, draw the toxicity of nano-particle；
Described nano-particle includes the one in nano ferriferrous oxide, nano titanium oxide, nano zine oxide, nano silicon, nano-Ag particles.
2. method according to claim 1, it is characterised in that in described step 1) deposition argent be shaped as square, the length of side range for 20 μm-100 μm.
3. method according to claim 1, it is characterised in that described step 2) in, when carrying out electrospinning, voltage is 15-25KV, and flow velocity is 0.5-1.0ml/h.
4. method according to claim 1, it is characterised in that described step 2) in, the diameter of fiber is 200nm.
5. method according to claim 1, it is characterised in that described step 2) in medical high polymer be polylactic acid, described organic solvent is the mixed solution of acetone and dimethylformamide, and the volume ratio of acetone and dimethylformamide is 9:1.
6. method according to claim 1, it is characterised in that the wide of the mould in described step 3) is 100 μm, and height is 80 μm.
7. method according to claim 1, it is characterised in that the wide of the mould in described step 3) is 50 μm, and height is 70 μm.
8. method according to claim 1, it is characterised in that in described step 5), the number of the Primary mouse myocardial cell for inoculating is 8 × 106Individual.
9. method according to claim 1, it is characterised in that in described step 5), the flow velocity of culture medium is 0.3ml/h.
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